Cell surface GRP78 regulates BACE2 via lysosome-dependent manner to maintain mesenchymal phenotype of glioma stem cells.
Cell surface GRP78 regulates BACE2 via lysosome-dependent manner to maintain mesenchymal phenotype of glioma stem cells.
复制标题
细胞表面GRP78通过溶酶体依赖性方式调节BACE2维持胶质瘤干细胞间充质表型
DOI：
10.1186/s13046-020-01807-4
复制
发表时间：
2021-01-07
期刊：
影响因子：
--
通讯作者：
Li G
中科院分区：
文献类型：
--
作者：
Chen Z;Wang H;Zhang Z;Xu J;Qi Y;Xue H;Gao Z;Zhao R;Wang S;Zhang S;Qiu W;Guo X;Li G
背景胶质瘤干细胞(GSCs)被认为是胶质瘤的起始细胞，对治疗产生抵抗作用。患者来源的GSCs很好地概括了其亲代胶质瘤组织的异质性，可分为不同的亚型。同样，以前的工作将GSCs分为两个不同的亚型，间充质(MES)和原神经(PN)亚型，普遍认为MES亚型被认为是一种更恶性的表型，具有高侵袭性和辐射抗性的特征。因此，了解MES表型的机制对于胶质母细胞瘤的治疗是必要的。方法从癌症基因组图谱(TCGA)和基因表达总览(GEO)数据库获得生物信息学分析数据。用抗体阻断细胞表面葡萄糖调节蛋白78(CsGRP78)。用细胞凋亡率和细胞周期分析来评估辐射损伤。免疫荧光染色检测蛋白质的表达和分布。采用质谱学和生物信息学相结合的方法筛选下游分子。结果GRP78的总表达与MES-GSC的干性有关，csGRP78在MES-GSC中高表达。靶向csGRP78抑制体外和体内MES GSCs的自我更新和辐射抗性，同时下调STAT3、NF-κB和C/EBPβ通路。质谱学显示潜在的下游β位点APP裂解酶2(BACE2)受csGRP78通过溶酶体降解调节。结论细胞表面GRP78在MES GSCs中优先表达，并在MES表型维持中起关键作用。因此，用高特异性抗体封闭MES GSCs中的csGRP78可能是一种有前景的新治疗策略。
BackgroundGlioma stem cells (GSCs) are considered the initial cells of gliomas, contributing to therapeutic resistance. Patient-derived GSCs well recapitulate the heterogeneity of their parent glioma tissues, which can be classified into different subtypes. Likewise, previous works identified GSCs as two distinct subtypes, mesenchymal (MES) and proneural (PN) subtypes, and with general recognition, the MES subtype is considered a more malignant phenotype characterized by high invasion and radioresistance. Therefore, understanding the mechanisms involved in the MES phenotype is necessary for glioblastoma treatment.MethodsData for bioinformatic analysis were obtained from The Cancer Genome Atlas (TCGA) and The Gene Expression Omnibus (GEO) database. An antibody was used to block cell surface glucose-regulated protein 78 (csGRP78). Apoptosis and cell cycle analyses were performed to evaluate radiation damage. Immunofluorescence staining was applied to assess protein expression and distribution. Mass spectrometry combined with bioinformatic analysis was used to screen downstream molecules. Intracranial GSC-derived xenografts were established for in vivo experiments.ResultsTotal GRP78 expression was associated with MES GSC stemness, and csGRP78 was highly expressed in MES GSCs. Targeting csGRP78 suppressed the self-renewal and radioresistance of MES GSCs in vitro and in vivo, accompanied by downregulation of the STAT3, NF-κB and C/EBPβ pathways. Mass spectrometry revealed the potential downstream β-site APP-cleaving enzyme 2 (BACE2), which was regulated by csGRP78 via lysosomal degradation. Knockdown of BACE2 inactivated NF-κB and C/EBPβ and significantly suppressed the tumorigenesis and radioresistance of MES GSCs in vitro and in vivo.ConclusionsCell surface GRP78 was preferentially expressed in MES GSCs and played a pivotal role in MES phenotype maintenance. Thus, blocking csGRP78 in MES GSCs with a high-specificity antibody might be a promising novel therapeutic strategy.
登录
查看更多内容
影响因子：
8.8
作者：
Mineo M;Ricklefs F;Rooj AK;Lyons SM;Ivanov P;Ansari KI;Nakano I;Chiocca EA;Godlewski J;Bronisz A
通讯作者：
Bronisz A
影响因子：
4.8
作者：
Gopal, Udhayakumar;Mowery, Yvonne;Pizzo, Salvatore Vincent
通讯作者：
Pizzo, Salvatore Vincent
影响因子：
50.3
作者：
Suva, Mario L.;Tirosh, Itay
通讯作者：
Tirosh, Itay
影响因子：
50.3
作者：
Phillips, HS;Kharbanda, S;Aldape, K
通讯作者：
Aldape, K
影响因子：
11.2
作者：
Galli, R;Binda, E;Vescovi, A
通讯作者：
Vescovi, A